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Effects of Desflurane and Sevoflurane anesthesia on regulatory T cells in patients undergoing living donor kidney transplantation: A randomized intervention trial

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Recognizing that regulatory T cells (Tregs) plays crucial roles in transplant tolerance and high peripheral blood Tregs associated with stable kidney graft function, knowing which volatile anesthetic agents can induce peripheral blood Tregs increment would have clinical implications. This study aimed to compare effects of desflurane and sevoflurane anesthesia on peripheral blood Tregs induction in patients undergoing living donor kidney transplantation.

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R E S E A R C H A R T I C L E Open Access

Effects of Desflurane and Sevoflurane

anesthesia on regulatory T cells in patients

undergoing living donor kidney

transplantation: a randomized intervention

trial

Arpa Chutipongtanate1, Sasichol Prukviwat1, Nutkridta Pongsakul2, Supanart Srisala3, Nakarin Kamanee1,

Nuttapon Arpornsujaritkun4, Goragoch Gesprasert4, Nopporn Apiwattanakul5, Suradej Hongeng6,

Wichai Ittichaikulthol1, Vasant Sumethkul7and Somchai Chutipongtanate2,8*

Abstract

Background: Volatile anesthetic agents used during surgery have immunomodulatory effects which could affect

postoperative outcomes Recognizing that regulatory T cells (Tregs) plays crucial roles in transplant tolerance and high peripheral blood Tregs associated with stable kidney graft function, knowing which volatile anesthetic agents can induce peripheral blood Tregs increment would have clinical implications This study aimed to compare effects of desflurane and sevoflurane anesthesia on peripheral blood Tregs induction in patients undergoing living donor kidney transplantation Methods: A prospective, randomized, double-blind trial in living donor kidney transplant recipients was conducted at a single center, tertiary-care, academic university hospital in Thailand during August 2015– June 2017 Sixty-six patients were assessed for eligibility and 40 patients who fulfilled the study requirement were equally randomized and allocated

to desflurane versus sevoflurane anesthesia during transplant surgery The primary outcome included absolute changes of peripheral blood CD4+CD25+FoxP3+Tregs which measured by flow cytometry and expressed as the percentage of the total population of CD4+T lymphocytes at pre-exposure (0-h) and post-exposure (2-h and 24-h) to anesthetic gas.P-value

< 0.05 denoted statistical significance

(Continued on next page)

© The Author(s) 2020 Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/ The Creative Commons Public Domain Dedication waiver ( http://creativecommons.org/publicdomain/zero/1.0/ ) applies to the

* Correspondence: schuti.rama@gmail.com ; somchai.chu@mahidol.edu

2

Pediatric Translational Research Unit, Department of Pediatrics, Faculty of

Medicine Ramathibodi Hospital, Mahidol University, Bangkok 10400, Thailand

8 Department of Clinical Epidemiology and Biostatistics, Faculty of Medicine

Ramathibodi Hospital, Mahidol University, Bangkok 10400, Thailand

Full list of author information is available at the end of the article

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(Continued from previous page)

Results: Demographic data were comparable between groups No statistical difference of peripheral blood Tregs

between desflurane and sevoflurane groups observed at the baseline pre-exposure (3.6 ± 0.4% vs 3.1 ± 0.4%;p = 0.371) and 2-h post-exposure (3.0 ± 0.3% vs 3.5 ± 0.4%;p = 0.319) At 24-h post-exposure, peripheral blood Tregs was

significantly higher in desflurane group (5.8 ± 0.5% vs 4.1 ± 0.3%;p = 0.008) Within group analysis showed patients

receiving desflurane, but not sevoflurane, had 2.7% increase in peripheral blood Treg over 24-h period (p < 0.001)

Conclusion: This study provides the clinical trial-based evidence that desflurane induced peripheral blood Tregs

increment after 24-h exposure, which could be beneficial in the context of kidney transplantation Mechanisms of action and clinical advantages of desflurane anesthesia based on Treg immunomodulation should be investigated in the future Trial registration: ClinicalTrials.gov,NCT02559297 Registered 22 September 2015 - retrospectively registered

Keywords: Clinical trial, Inhalation agent, Kidney transplant, Tregs, Volatile anesthesia

Background

Kidney transplantation is the best option for renal

replace-ment therapy in patients with end-stage renal disease

(ESRD), often restoring quality of life in ESRD patients

Allograft rejection, an immune-mediated process, is a

common cause of transplant failure [1–4] Evidence

indi-cates CD4+CD25+FoxP3+cells, commonly known as

regu-latory T cells (Tregs), play a critical role in preventing

graft rejection by suppression of recipient alloimmune

re-sponse [5–8] In healthy subjects, Tregs represent up to

5% of peripheral CD4+T cells [9–11] In kidney transplant

patients, high peripheral blood Tregs were associated with

stable graft function Low peripheral blood Tregs was

as-sociated with allograft rejection [12–17]

Currently, adoptive transfer of ex vivo expanded Tregs

is a promising strategy to induce transplant tolerance and

control graft rejection in kidney transplant recipients [18,

19] It has been investigated for safety and feasibility in

phase I trials, i.e., the ONE (NCT02091232) and TRACT

(NCT02145325) Identifying Treg-friendly agents from

pharmacologic choices in multiple steps of kidney

trans-plant management may also offer an attractive therapeutic

strategy [19] Characterization of Tregs under various

treatment conditions may help refine current preventive

measures or identify novel therapeutic targets

Volatile anesthetic agents are widely used for general

anesthesia during kidney transplantation A growing

body of evidence from ex vivo and clinical studies [20–

26] suggest desflurane and sevoflurane (halogenated

ether inhaled agents) exhibit immunomodulatory effects

(e.g., cell proliferation, activation, migration, cytokine

production) on neutrophils, macrophages, natural killer

cells, B and T lymphocytes These effects may be

medi-ated via activation of volatile anesthetic receptors (i.e.,

γ-aminobutyric acid type A receptor, nicotinic

acetylcho-line receptor, serotonin receptor and non-canonical

β2-integrins) or via binding to surface adhesion molecules

such as integrin leukocyte function associated antigen-1,

which express differentially on peripheral blood

leuko-cytes [25, 26] However, effects of desflurane and

sevoflurane on Treg immunomodulation is surprisingly overlooked and has only rarely been investigated A bet-ter understanding of these effects would have transla-tional potential For example, the early Treg immunomodulation by anesthetic agents may help miti-gating the initiation of alloimmune responses during the 24-h perioperative period, and works in conjunction with the standard immunosuppressive regimen to seam-lessly maintain the graft survival in LDKT patients This interventional trial aims to compare the immuno-modulatory effects of desflurane and sevoflurane anesthesia on peripheral blood Treg induction in pa-tients undergoing living donor kidney transplantation (LDKT) Several plasma cytokines were measured as the surrogate outcomes of the volatile anesthetic effects on anti- and pro-inflammatory responses Evidence from this study would support future investigation of volatile anesthetic agents as part of perioperative management with an aim to improve transplant outcomes

Methods

Trial design and patient enrollment

This prospective, double-blind, randomized interven-tion trial was approved by the Ethical Clearance Committee on Human Rights Related to Research In-volving Human Subjects, Faculty of Medicine Ramathibodi Hospital, Mahidol University (protocol

ID 045823) and the protocol was registered to Clini-calTrials.gov (identifier NCT02559297) on September

22, 2015 Patients aged ≥18 years old who received their first living donor kidney transplantation at Ramathibodi Hospital were included in the study Pa-tients were excluded for hyperacute graft rejection, currently on immunosuppressive drugs due to under-lying diseases, receiving blood products during 24-h perioperative period, or patient refusal to participate

in the study at any time point Informed consent was obtained from all subjects No interim analysis was performed during the trial This study followed the CONSORT reporting guideline [27]

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Randomization was generated in a 1:1 allocation with a

block size of 8 and the random number was put in a

sealed envelope Patients were randomly assigned to

ei-ther desflurane or sevoflurane intervention by drawing a

sealed envelope Randomization took place on the day of

surgery just prior to initiation of anesthesia

Blinding

Subjects and outcome assessors (including laboratory

tech-nicians and all investigators except the designated research

coordinator) were blinded to group allocation throughout

surgery, laboratory investigation and data collection

Blind-ing was uncovered at the time of data analysis

Interventions

Patients were randomly assigned to receive desflurane or

sevoflurane for the maintenance phase of anesthesia In

addition to the randomized inhalation agents, patients

received the same regimen of 1–2 mg of midazolam for

premedication and intravenous anesthetic agents

includ-ing 1–2 mcg kg− 1of fentanyl, 1–2 mg kg− 1of propofol

and 0.5–0.6 mg kg − 1 of atracurium for induction of

anesthesia and intubation A balance anesthesia

tech-nique was used for maintenance phase The inhalation

agent (sevoflurane or desflurane) was used in

conjunc-tion with 50% nitrous oxide in oxygen Ventilaconjunc-tion was

adjusted to maintain normocarbia End-tidal anesthetic

gas monitoring was used to ensure 1.0–1.5 minimum

al-veolar concentration (MAC) of the inhalation agent

dur-ing maintenance phase in both groups

During anesthesia, blood pressure, heart rate, oxygen

saturation, ETCO2, and temperature were monitored

and recorded Blood pressure was maintained within

20% of baseline values Hypotension was managed by

intravenous fluid and ephedrine IV bolus as needed

Total doses of intravenous medications were recorded

All patients in both interventions were transferred to the

kidney transplant unit for postoperative care

Blood sample collection

Venipuncture was performed at three time-points;

pre-exposure (0-h) and post-pre-exposure (2-h, and 24-h) to

in-halation agents Two tubes of 0.5-ml EDTA blood were

collected at each time point, one for Treg enumeration

and the other for cytokine measurement

Outcome measures

The primary outcome was the absolute change in

num-ber of peripheral blood CD4+CD25+FoxP3+Tregs, which

was measured by flow cytometry and expressed as the

percentage of the total population of CD4+ T

lympho-cytes at pre-exposure (0-h) and post-exposure (2-h and

24-h) to anesthetic gas A secondary outcome was the

plasma level of anti-inflammatory cytokine IL-10 (the major cytokine produced by Tregs), TGF-β1 (anti-in-flammatory cytokines produced by many types of cells and required for Tregs differentiation), and pro-inflammatory cytokines produced by T helper (Th) 1/ Th2, i.e., GM-CSF, IFN-ɣ, IL-2, IL-4, IL-5, IL-12, IL-13 and TNF-α, which measured by multiplex immunoassay All measurements were performed in triplicate

Treg enumeration by flow cytometry

Peripheral blood mononuclear cells (PBMC) were iso-lated by density gradient centrifugation Approximately

5 × 105cells were suspended in 20μL phosphate buffer saline (PBS) in the presence of cell surface marker anti-bodies (APC-CD4, PE-Cy7-CD25) (#MHCD0404, #25– 0259-41; ThermoFisher, Florence, KY), mixed well and incubated at room temperature for 15 min Thereafter, cells were permeabilized and intracellularly stained using FoxP3-FITC antibody (#11–4776-42; ThermoFisher) Flow cytometry (BD FACSVerse with BD FACSuite soft-ware; BD Bioscience, San Jose, CA) was used to measure the number of Tregs, expressed as a percentage of CD4+CD25+FoxP3+T cells among the CD4+cell popula-tion The estimated number of CD4 + cells and Treg were calculated by determining the ratio of CD4 + cell count and CD4 + CD25 + FoxP3 + cell count, respect-ively, to the total count in the flow cytometry, and then multiplied by the number of white blood cells measured from the complete blood count (CBC) which ordered at the pre-operative and post-operative evaluations

Cytokine measurement by multiplex immunoassay

Multiplex cytokine immunoassay was performed by BioPlex-200 system (Bio-Rad, Hercules, CA) GM-CSF, IFN-ɣ, IL-2, IL-4, IL-5, IL-10, IL-12, IL13, and TNF-α were detected by BioPlex Pro human cytokine Th1/Th2 assay (Bio-Rad), and TGF-β1 was measured by single-plex custom assay (Bio-Rad) as the manufacturer’s instruction

Sample size estimation and statistical analysis

There was no data related to sevoflurane and desflurane anesthesia on Treg immunomodulation available at the initiation of the study Nevertheless, Pirbudak Cocelli L

et al [21], showed that sevoflurane and desflurane anesthesia caused a significant difference in total lymphocyte count at 2-h post-induction in patients undergoing abdominal surgery Since Treg is a subset of lymphocytes, our study then adopted mean difference and standard deviation to calculate the effect size The nQuery Advisor program was applied for sample size calculation Accordingly, at least 40 patients (20 patients per group) were required to determine statistically sig-nificant mean difference between groups (the effect size

of 0.915, alpha = 0.05 and power = 80%)

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Statistical analysis was performed by Excel and R

pack-ages Data were reported in number, percentage, mean ±

SD (or SEM) or median [IQR] as appropriate Parametric

and non-parametric tests were used, as appropriate, to

determine difference between groups ANOVA with

Tukey post-hoc test was performed for multiple

com-parison P-value < 0.05 was considered to be statistically

significant

Results

Baseline characteristics

Figure1shows the flow of the participants in this study

A total of 66 patients were assessed for eligibility and 46

patients who met inclusion criteria were recruited

dur-ing August 10, 2015 to June 3, 2017 for randomization

and allocation to the intervention Six patients who met

exclusion criteria after allocation due to receiving

peri-operative blood products were excluded Table1 shows

demographic and clinical data for the 40 patients

en-rolled in this study Most variables, including recipient

factors, donor factors, protocol immunosuppressive

regi-mens, intraoperative parameters, the dosage of

intraven-ous anesthesia were not significantly different between

the intervention groups Donor age and the estimated

blood loss were slightly lower in patients receiving

des-flurane anesthesia Factors contributing to

ischemic-reperfusion injury, i.e., cold and warm ischemic time, were comparable between groups Also, anesthesia time was not different between groups (279 ± 42 min vs

303 ± 45 min, p = 0.098) Given that the depth of sevo-flurane and dessevo-flurane anesthesia was maintained at 1.0–1.5 MAC for each arm, this finding supported that patients were exposed to inhalation agents equally (Table1)

Effects of Desflurane and Sevoflurane anesthesia on Tregs

in LDKT recipients

Figure 2a demonstrates the gating strategy of flow cy-tometry and Fig.2b shows the effects of sevoflurane and desflurane anesthesia on CD4+CD25+FoxP3+ Tregs in peripheral blood of LDKT recipients (n = 20 per group)

No significant difference of peripheral blood Tregs (mean ± SEM) was observed at pre-exposure (3.6 ± 0.4%

vs 3.1 ± 0.4%; p = 0.371) and 2-h post-exposure (3.0 ± 0.3% vs 3.5 ± 0.4%; p = 0.319) between desflurane and sevoflurane, respectively However, at 24-h post-exposure, desflurane group had significantly higher per-ipheral blood Tregs as compared to sevoflurane group (5.8 ± 0.5% vs 4.1 ± 0.3%; p = 0.008) (Fig 2b) Within-group analysis showed that the patients receiving des-flurane had 2.7% increase in Tregs over 24-h period (p <

Fig 1 Flow diagram of study participants

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0.001) (Fig 2b), while this effect was not observed in

sevoflurane group

Although peripheral blood Treg are commonly presented

in the literatures as the percentage of CD25+FoxP3+cells in

the CD4+cell population, one argument was that the

incre-ment in Treg percentage might be corresponding to the

global changes of leukocytes or CD4+T cells in response of surgical procedure and postoperative inflammation but not the influence of inhalation agents To address this issue, the absolute number of white blood cells (as measured by the complete blood count), and the calculated numbers of CD4+ T cells and CD4+CD25+FoxP3+Tregs (details in the

Table 1 Demographic data of LDKT recipients enrolled into the study If not indicated otherwise: n (%)

Recipient

Donor

Protocol immunosuppressive drug

Intraoperative variable

Intravenous anesthesia

Abbreviations: ADPKD Autosomal dominant polycystic kidney disease, BMI Body mass index, HLA Human leukocyte antigen, PRA Panel reactive antibodies

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Method section), were shown in Fig 2c Postoperative

leukocytosis was observed in both groups as expected (Fig

2c, the left panel), while the number of CD4+ T cells were

not significantly changed during 24-h perioperative period

(Fig 2c, the center panel) Consistently, the number of

Tregs (median [IQR]) were comparable between groups at

the baseline (7 [6,11] vs 8 [4,9] cells/mm3;p = 0.780) and

were significantly higher in desflurane group (12 [9,16] vs

8 [4,12] cells/mm3;p = 0.033) at 24-h post-exposure (Fig

2c, the right panel) This finding suggested the effect of

des-flurane anesthesia on the peripheral blood Treg induction

during 24-h postoperative period

Plasma cytokine levels were measured by multiplex

im-munoassay as a surrogate outcome of immunomodulation

possibly influenced by inhalation agents (n = 26, 12

sevo-flurane and 14 dessevo-flurane) Although there was no

statisti-cally significant difference between groups in any

cytokine, a trend of increased IL-10 was observed in

des-flurane group as compared to sevodes-flurane group at 24-h

post-exposure (27.5 [17.6, 34.4] vs 17.8 [11.4, 22.3] pg/

mL;p = 0.12) (Fig 3 and Supplementary Table1) IL-10,

the signature anti-inflammatory cytokine produced by

Tregs, had an upward trend (1.67-time increased at 24-h

as compared to the baseline pre-exposure) in the patients

receiving desflurane, whereas other cytokines seemed to

be unchanged over the 24-h period (Fig 3 and Supple-mentary Table 1) The transient drop of measured cyto-kines at 2-h was potentially associated with intraoperative factors, e.g., intravenous fluid administration, but not dir-ectly influenced by inhalation agents

An increased trend of plasma IL-10 in the desflurane group was in line with previous results (Fig 2) and sug-gested that desflurane anesthesia was associated with IL-10-producing Tregs induction in LDKT patients during the perioperative period Matched-pair data of Tregs and plasma IL-10 levels in 26 patients (14 desflurane, 12 sevoflurane) were analyzed to observe this immunophenotypic response Scatter plot showed a positive relationship between Tregs and IL-10 fold changes over 24-h period (Fig.4a), in which the proportion of patients with increased Tregs and IL-10 immunophenotypic response was higher in the desflurane group (Fig 4b) Taken together, our findings revealed that desflurane anesthesia induced IL-10-producing Tregs in LDKT recipients over 24-h postoperative period

Discussion

Increasing evidence suggests that volatile anesthetic agents exhibit immunomodulatory effects linked to innate and

Fig 2 Effects of desflurane and sevoflurane anesthesia on the CD4 + CD25 + FoxP3+ Tregs ( n = 20 per group) a Representative gating strategy of Treg enumeration b The percentage of Treg in the CD4 + cell population c The absolute number of white blood cells (left), and the calculated numbers of CD4 + cells (center) and Treg (right), in peripheral blood (details in the Method section) The result showed that the LDKT patients who received desflurane anesthesia, but not sevoflurane, had significantly increased Tregs in the peripheral blood at 24-h post-exposure All experiments were performed in triplicate Des, desflurane; NS, not significant; Sevo, sevoflurane

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adaptive immunity via induction and suppression of

neu-trophils, macrophages, NK cells and B and T lymphocytes

[25,26] However, their effects on Treg have remained

un-known This study, for the first time, showed that

desflur-ane, but not sevoflurdesflur-ane, increased Treg frequency in

peripheral blood of LDKT recipients during 24-h

peri-operative period Selection of desflurane anesthesia in

kid-ney transplantation may have additional benefits to kidkid-ney

graft outcome, particularly preventing allograft rejection

Studies showed that kidney transplant patients who

maintained a high level of peripheral blood Tregs were

associated with better outcomes [14, 15, 28] San

Segundo D, et al [14], reported that among 90 kidney

transplant recipients, patients who maintained high

levels (above 70th percentile) of peripheral blood Tregs

at both 6 and 12 months had a better prognosis in the

aspect of long-term graft survival after 4 and 5 years

follow-up Liu L, et al [15], compared peripheral blood

Treg levels between 42 patients with stable kidney graft

function and ten patients who suffered from chronic

rejection The results showed that Treg levels were sig-nificantly higher in the stable group than the chronic re-jection group Alberu J, et al [28], investigated the association between Treg levels and de novo donor-specific HLA-antibody (DSA) production in 53 kidney transplant patients Although early development of DSA was not associated with Treg numbers, at 12 months after kidney transplant DSA-negative patients had higher number of peripheral blood Treg

The mechanisms for which higher peripheral blood Tregs help prevention of allograft rejection and mainten-ance of transplant tolermainten-ance meet the same concept of peripheral regulation in autoimmune reaction [18, 29–

32] On a cellular basis, Tregs utilize four modes of ac-tion in peripheral regulaac-tion including [29–32]; i) secre-tion or generasecre-tion of inhibitory cytokines (e.g., 10,

IL-35, TGF-β and adenosine); ii) direct killing of targets through Granzyme A/B and perforin-dependent cytoly-sis; iii) IL-2 consumption through high IL-2R expression which leads to cytokine-mediated deprivation and

Fig 3 Plasma cytokines were measured by multiplex cytokine immunoassay Box plots exhibited plasma levels of anti-inflammatory cytokines

IL-10 and TGF- β1, and pro-inflammatory cytokines GM-CSF, IFN-γ, IL-2, IL-4, IL-5, IL-12, IL-13 and TNF-α (n = 26; 14 desflurane, 12 sevoflurane) IL-10 showed an increased trend over 24-h period in patients receiving desflurane anesthesia Des, desflurane; Sevo, sevoflurane

Fig 4 Matched-pair data analysis of Tregs and IL-10 ( n = 26; 14 desflurane, 12 sevoflurane) Fold change was calculated by Tregs (or IL-10) measured at 24-h divided by that of pre-exposure (0-h) in the same patient, in which fold change > 1 indicated upregulation and fold change <

1 was downregulation a Scatter plot exhibited the positive relationship between Tregs and IL-10 fold changes b Bar plots showed a higher proportion of co-increased Tregs and IL-10 immunophenotypic response in patients receiving desflurane anesthesia Des, desflurane;

Sevo, sevoflurane

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apoptosis of effector cells; and iv) direct interaction with

CTLA-4, LAG-3 and PD1 molecules Although

accumu-lating evidence would favor contact-dependent

mecha-nisms over non-contact/secretory component alone,

these different mechanisms should work in concert to

control various immune effector cells and regulate

dif-ferent inflammatory settings [29–32] Given that breadth

of regulatory function on autoimmunity and

self-tolerance, changes in peripheral blood Tregs in kidney

transplant recipients may shift the balance between

allo-graft rejection and transplant tolerance

According to these lines of evidence, the increment of

Tregs and IL-10 (Figs.2,3 and4) after exposure to

des-flurane anesthesia should be beneficial to graft outcomes

in LDKT recipients In fact, several drugs routinely used

in general anesthesia (besides volatile anesthetic agents)

have immunomodulatory properties [25, 26] It is

pos-sible that some of them have positive effects on Tregs

Synergistic effects of multiple Treg-modulated agents

may provide a better transplant outcome Understanding

how anesthetic agents exhibit varied effects on the

im-mune system, particularly on Tregs, is important for

fu-ture development of perioperative medicine in kidney

transplantation

This study was associated with several limitations

First, clinical outcomes, such as short-term and

long-term graft survival, that were associated with Treg

immunomodulation of desflurane anesthesia were not

investigated Graft survival is influenced by various

fac-tors (such as adequacy and toxicity of

immunosuppres-sive drugs, presence of donor-specific antigen, PRA

levels, numbers of HLA mismatch) These factors,

to-gether with Treg immunomodulation of desflurane,

should be taken into account in future studies Secondly,

plasma cytokine levels were measured in 26 out of 40

patients (2/3 of total population in this study) due to the

limited budget A non-significant difference of cytokines

between intervention groups may be due to a lack of

statistical power, but at least, the upward trend of

plasma IL-10 was observed, given the supportive

evi-dence that desflurane anesthesia induced peripheral

blood Tregs with potential IL-10 production Third, the

mechanisms of action (MoA) that drive desflurane-Treg

immunomodulation were not defined and not the focus

of this study Further studies to characterize receptors

and downstream signaling pathways that are responsible

for desflurane-Treg effects would give some insight into

a new MoA class of volatile anesthetic agent or a new

biological process that facilitates transplant tolerance in

LDKT recipients

Conclusion

In summary, desflurane had the advantage over

sevoflur-ane as the inhalation sevoflur-anesthetic in LDKT patients

regarding the increment of peripheral blood Tregs Fur-ther research focused on clinical outcomes and pharma-cological actions of desflurane on Treg immunomodulation has translational potential, which could eventually benefit LDKT recipients

Supplementary information

Supplementary information accompanies this paper at https://doi.org/10 1186/s12871-020-01130-7

Additional file 1: Table S1 Effects of sevoflurane and desflurane anesthesia on plasma cytokine levels of LDKT patients.

Abbreviations

ADPKD: Autosomal dominant polycystic kidney disease; BMI: Body mass index; ESRD: End-stage renal disease; HLA: Human leukocyte antigen; LDKT: Living donor kidney transplantation; MoA: Mechanisms of action; PRA: Panel reactive antibodies; Th1: T helper 1; Th2: T helper 2;

Tregs: Regulatory T cells

Acknowledgments

We are grateful to all staff of the Ramathibodi Kidney Transplant Project SC was financially supported by Faculty Staff Development Program of Faculty

of Medicine Ramathibodi Hospital, Mahidol University, for his research activities.

Authors ’ contributions

AC and SC initiated the conception AC, NOA, GG, WI, VS, SC developed the design SP performed patient recruitment, randomization, and allocation AC and WI performed anesthesia intervention NUA, GG, VS provided medical care and surgical operation NP, SS, NK, NOA, SH, SC performed Treg enumeration and cytokine measurement AC, SP, SC analyzed data, prepared figures and tables AC and SP wrote the manuscript NP, SS, NK, NOA, GG, NUA, SH, WI, VS, SC revised the manuscript All authors read and approved the final version to be published.

Funding This study was supported by Talent Management Program of Mahidol University, Thailand (TM:CP131 to AC) The funder had no role in study design, collection, and analysis of data and the decision to publish the manuscript.

Availability of data and materials The datasets used and/or analyzed during the current study are available from the corresponding author on reasonable request.

Ethics approval and consent to participate This study was approved by the Ethical Clearance Committee on Human Rights Related to Research Involving Human Subjects, Faculty of Medicine Ramathibodi Hospital, Mahidol University (protocol ID 045823) and the protocol was registered to ClinicalTrials.gov (identifier NCT02559297) All patients provided their written informed consents to participate in this study.

Consent for publication Not applicable.

Competing interests The authors declare no conflict of interests.

Author details

1 Department of Anesthesiology, Faculty of Medicine Ramathibodi Hospital, Mahidol University, Bangkok 10400, Thailand 2 Pediatric Translational Research Unit, Department of Pediatrics, Faculty of Medicine Ramathibodi Hospital, Mahidol University, Bangkok 10400, Thailand 3 Research Center, Faculty of Medicine Ramathibodi Hospital, Mahidol University, Bangkok

10400, Thailand 4 Vascular and Transplantation Unit, Department of Surgery, Faculty of Medicine Ramathibodi Hospital, Mahidol University, Bangkok

10400, Thailand 5 Division of Infectious Disease, Department of Pediatrics,

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Faculty of Medicine Ramathibodi Hospital, Mahidol University, Bangkok

10400, Thailand 6 Division of Hematology and Oncology, Department of

Pediatrics, Faculty of Medicine Ramathibodi Hospital, Mahidol University,

Bangkok 10400, Thailand.7Division of Nephrology, Department of Medicine,

Faculty of Medicine Ramathibodi Hospital, Mahidol University, Bangkok

10400, Thailand 8 Department of Clinical Epidemiology and Biostatistics,

Faculty of Medicine Ramathibodi Hospital, Mahidol University, Bangkok

10400, Thailand.

Received: 20 June 2020 Accepted: 20 August 2020

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